使用双光子激光共聚焦显微镜对活体小鼠脑组织(体感皮层)进行形态学和细胞内Ca^2+变化水平的观察和记录。方法是对小鼠开颅后向脑内注射Ca^2+敏感荧光染料OregonGreen488BAVTA-1乙酰氧基甲酯,然后在双光子显微镜下观察并记录。双光子显微镜下的图像清晰生动,可扫描到大脑皮层以下较深区域,并能检测神经元细胞内Ca^2+离子即时水平变化。实验结果表明通过注射Ca^2+敏感荧光染料并结合多光子激光共聚焦显微镜观察活体动物脑部变化的方法可以得到高分辨率图像并且作为分析的来源。
To observe the brain image and the change of Ca^2+ level in live mouse by using two-photon laser scanning confocal microscope. Using craniotomy to expose target area of mouse brain ( somatosensory cortex) then inject Ca^2+ sensitive fluorescent indicator, for instance, Oregon Green 488 BAPTA -1 acetoxymethyl. After that, depose the mouse under the two-photon to observe and record. The brain image record by two-photon is clear and vivid . And photograph indicate that by using two-photon confocal can reach deeper area of the brain and record the fast change of Ca^2+ in single neuron. The results show that by using multi-photon confocal microscope couple with calcium sensitive fluorescent indicator can get high resolution image of brain of live mouse.